LOCUS       40924_6252        6495 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Integrational vector pBEXK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6495)
  AUTHORS   Nikel PI, de Lorenzo V.
  TITLE     Implantation of unmarked regulatory and metabolic modules in 
            Gram-negative bacteria with specialised mini-transposon delivery 
            vectors
  JOURNAL   J. Biotechnol. 163 (2), 143-154 (2013)
  PUBMED    22609234
REFERENCE   2  (bases 1 to 6495)
  AUTHORS   Nikel PI, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-FEB-2012) Systems Biology Program, Centro Nacional de 
            Biotecnologia (CNB-CSIC), C/Darwin 3, Madrid, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 6495)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6495)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol."; date: "2013"; volume: "163"; issue: "2"; pages: 
            "143-154"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-FEB-2012) Systems Biology Program, Centro Nacional de 
            Biotecnologia (CNB-CSIC), C/Darwin 3, Madrid, Madrid 28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..19
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(41..1003)
                     /codon_start=1
                     /label=XylS
                     /note="XylS regulator encoded by the Pseudomonas putida TOL
                     plasmid pWWO"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     promoter        1903..1983
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     misc_feature    2033..2089
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      2094..2188
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    2199..2246
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2260..3072)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    3174..3221
                     /label=FRT
                     /note="FRT"
     protein_bind    3174..3221
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    complement(3228..3246)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             3340..4767
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     promoter        4824..4915
                     /label=AmpR promoter
     CDS             4916..5773
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      5780..5819
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            5966..6074
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      6093..6481
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctgtctctta tacacatctt taattaaacg ttcgtaatca agccacttcc tttttgcatt
       61 gacgcagggt gtcggaaggc aactcgccga acgcgctcct atagttttca gcgaagcgtc
      121 ccaaatgtaa gaagccgtag tctagggcta tctcagttat actacgcaca ttggcactgg
      181 gatcgttcaa gcaggcgcgg atgctttcga gcttgcggtt gcggatgtag ttcttcggcg
      241 tggtgccggc gtgcttctcg aacaaattgt agagcgagcg tggactcatc atcgccagct
      301 ccgctaaccg ctcaaggctg atattccgtt tgagattctc ctcaatgaat tgaacgactc
      361 gctcgaaaga cgggttacct ttgctgaaaa tttcacggct gacattgctg cccagcattt
      421 cgagcagctt ggaagcgatg atccccgcat agtgctcttg gacccgaggc atcgactttg
      481 tatgttccgc ttcgtcacaa actaacccga gtagattgat aaagccatcg agttgctgga
      541 gattgtgtcg cgcggcgaaa cggataccct ccctcggctt gtgccaattg ttgtcactgc
      601 acgcccgatc aaggaccact gagggcaatt taacgataaa tttctcgcaa tcttctgaat
      661 aggtcaggtc ggcttggtca tccggattga gcagcaatag ttcgcccggc gcaaaatagt
      721 gctcctggcc atggccacgc cacaggcaat ggcctttgag tattatttgc agatgataac
      781 aggtctctaa tccaggcgag attaccctca cgctaccgcc gtagctgatt cgacacagat
      841 cgaggcatcc gaagattctg tggtgcagcc tgcctgccgg gcgcccgccc ttgggcaggc
      901 gaatagagtg cgtaccgaca tactggttaa cataatcgga gactgcatag ggctcggcgt
      961 ggacgaagat ctgacttttc tcgttcaata agcaaaaatc catagttcac ggttctctta
     1021 ttttaatgtg ggctgcttgg tgtgatgtag aaaggcgcca agtcgatgaa aatgcatctc
     1081 gacgtgatgc gtatacgggt tacccccatt gccacgttgc gccatccttt ttgcaatcag
     1141 tgaccacttt tccaagcaaa aataacgcca agcagaacga agacgttctt tttaagaagc
     1201 gagaacacca gaagttcgtg ctgtcggggc atggggcgac gaattggcgg ataaagggga
     1261 tctgctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     1321 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccgtaattac gtatggcaat
     1381 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     1441 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     1501 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     1561 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     1621 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgc atgggaatta
     1681 gcttgatctg accaacgacc ggtagcggag ctatccaacg gcggtatacc aggaaaacac
     1741 acagcaggta catcagaaca gtaccatgac tgaagaacaa atagtttttt cctgatccat
     1801 aaagcagaac ggcctgctcc atgacaaatc tggctcccca actaatgccc catgcagcca
     1861 gcataaccag cataaagtgc agtgtccggt ttgataggga taagtccagc cttgcaagaa
     1921 gcggatacag gagtgcaaaa aatggctatc tctagtaagg cctacccctt aggctttatg
     1981 caacagaaac aataataatg gagtcatgac catgcctagg ccgcggccgc gcgaattcga
     2041 gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttc ttggactcct
     2101 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     2161 ccgccgggcg ttttttattg gtgagaatcc agactagtga agttcctatt ccgaagttcc
     2221 tattctctag aaagtatagg aacttccaat taattattag aaaaattcat ccagcatcag
     2281 atgaaattgc agtttgttca tatccggatt atcaatgcca tatttctgaa acagacgttt
     2341 ttgcaggctc gggctaaatt cgcccaggca gttccacaga atggccagat cctgataacg
     2401 atccgcaatg cccacacggc ccacatcaat gcagccaatc agtttgcctt catcgaaaat
     2461 caggttatcc aggctaaaat cgccgtgggt caccacgcta tccgggctaa acggcagcag
     2521 tttatgcatt tctttccaca cctgttccac cggccagccg ttacgttcat catcaaaatc
     2581 gctcgcatcc accaggccgt tgttcatacg gctctgcgcc tgggccagac gaaacacacg
     2641 atcgctgtta aacgggcagt tgcacaccgg aatgctatgc agacgacgca gaaacacggc
     2701 cagcgcatcc acaatgtttt cgccgctatc cggatattct tccagcacct gaaacgcggt
     2761 tttgcccgga atcgcggtgg tcagcagcca cgcatcatcc ggggtgcgaa taaaatgttt
     2821 aatggtcggc agcggcataa attcggtcag ccagttcaga cgcaccattt catcggtcac
     2881 atcgttcgcc acgctgcctt tgccatgttt cagaaacagt tccggcgcat ccggtttgcc
     2941 atacagacga taaatggtcg cgccgctctg acccacgtta tcacgcgccc atttatagcc
     3001 atacagatcc gcatccatgt tgctgttcag acgcggacgg ctacagctcg tttcacgctg
     3061 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     3121 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caagaagttc
     3181 ctattccgaa gttcctattc tctagaaagt ataggaactt cgacgtcaga tgtgtataag
     3241 agacagctgg tttaaactac acaagtagcg tcctgaacgg aacctttccc gttttccaga
     3301 atctgatgtt ccatgtgacc tcctaacatg gtaacgttca tgattaccag tgcactgcat
     3361 cgtgcggcgg attgggcgaa aagcgtgttt tctagtgctg cgctgggtga tccgcgtcgt
     3421 accgcgcgtc tggtgaatgt tgcggcgcaa ctggccaaat atagcggcaa aagcattacc
     3481 attagcagcg aaggcagcaa agccatgcag gaaggcgcgt atcgttttat tcgtaatccg
     3541 aacgtgagcg cggaagcgat tcgtaaagcg ggtgccatgc agaccgtgaa actggcccag
     3601 gaatttccgg aactgctggc aattgaagat accacctctc tgagctatcg tcatcaggtg
     3661 gcggaagaac tgggcaaact gggtagcatt caggataaaa gccgtggttg gtgggtgcat
     3721 agcgtgctgc tgctggaagc gaccaccttt cgtaccgtgg gcctgctgca tcaagaatgg
     3781 tggatgcgtc cggatgatcc ggcggatgcg gatgaaaaag aaagcggcaa atggctggcc
     3841 gctgctgcaa cttcgcgtct gagaatgggc agcatgatga gcaacgtgat tgcggtgtgc
     3901 gatcgtgaag cggatattca tgcgtatctg caagataaac tggcccataa cgaacgtttt
     3961 gtggtgcgta gcaaacatcc gcgtaaagat gtggaaagcg gcctgtatct gtatgatcac
     4021 ctgaaaaacc agccggaact gggcggctat cagattagca ttccgcagaa aggcgtggtg
     4081 gataaacgtg gcaaacgtaa aaaccgtccg gcgcgtaaag cgagcctgag cctgcgtagc
     4141 ggccgtatta ccctgaaaca gggcaacatt accctgaacg cggtgctggc cgaagaaatt
     4201 aatccgccga aaggcgaaac cccgctgaaa tggctgctgc tgaccagcga gccggtggaa
     4261 agtctggccc aagcgctgcg tgtgattgat atttataccc atcgttggcg cattgaagaa
     4321 tttcacaaag cgtggaaaac gggtgcgggt gcggaacgtc agcgtatgga agaaccggat
     4381 aacctggaac gtatggtgag cattctgagc tttgtggcgg tgcgtctgct gcaactgcgt
     4441 gaatctttta ctccgccgca agcactgcgt gcgcagggcc tgctgaaaga agcggaacac
     4501 gttgaaagcc agagcgcgga aaccgtgctg accccggatg aatgccaact gctgggctat
     4561 ctggataaag gcaaacgcaa acgcaaagaa aaagcgggca gcctgcaatg ggcgtatatg
     4621 gcgattgcgc gtctgggcgg ctttatggat agcaaacgta ccggcattgc gagctggggt
     4681 gcgctgtggg aaggttggga agcgctgcaa agcaaactgg atggctttct ggccgcgaaa
     4741 gacctgatgg cgcagggcat taaaatctaa tcgatttaaa ttagtagccc gcctaatgag
     4801 cgggcttttt tttaattccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4861 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4921 cattcagcat tttcgtgtgg cgctgattcc gttttttgcg gcgttttgcc tgccggtgtt
     4981 tgcgcatccg gaaaccctgg tgaaagtgaa agatgcggaa gatcaactgg gtgcgcgcgt
     5041 gggctatatt gaactggatc tgaacagcgg caaaattctg gaatcttttc gtccggaaga
     5101 acgttttccg atgatgagca cctttaaagt gctgctgtgc ggtgcggttc tgagccgtgt
     5161 ggatgcgggc caggaacaac tgggccgtcg tattcattat agccagaacg atctggtgga
     5221 atatagcccg gtgaccgaaa aacatctgac cgatggcatg accgtgcgtg aactgtgcag
     5281 cgcggcgatt accatgagcg ataacaccgc ggcgaacctg ctgctgacga ccattggcgg
     5341 tccgaaagaa ctgaccgcgt ttctgcataa catgggcgat catgtgaccc gtctggatcg
     5401 ttgggaaccg gaactgaacg aagcgattcc gaacgatgaa cgtgatacca ccatgccggc
     5461 agcaatggcg accaccctgc gtaaactgct gacgggtgag ctgctgaccc tggcaagccg
     5521 ccagcaactg attgattgga tggaagcgga taaagtggcg ggtccgctgc tgcgtagcgc
     5581 gctgccggct ggctggttta ttgcggataa aagcggtgcg ggcgaacgtg gcagccgtgg
     5641 cattattgcg gcgctgggcc cggatggtaa accgagccgt attgtggtga tttataccac
     5701 cggcagccag gcgacgatgg atgaacgtaa ccgtcagatt gcggaaattg gcgcgagcct
     5761 gattaaacat tggtaaaccg atacaattaa aggctccttt tggagccttt ttttttggac
     5821 gacccttgtc cttttccgct gcataaccct gcttcggggt cattatagcg attttttcgg
     5881 tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc
     5941 cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg
     6001 ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc
     6061 tgcgaggctg gccgtaggcc ggccggcgcg ccgatctgaa gatcagcagt tcaacctgtt
     6121 gatagtacgt actaagctct catgtttcac gtactaagct ctcatgttta acgtactaag
     6181 ctctcatgtt taacgaacta aaccctcatg gctaacgtac taagctctca tggctaacgt
     6241 actaagctct catgtttcac gtactaagct ctcatgtttg aacaataaaa ttaatataaa
     6301 tcagcaactt aaatagcctc taaggtttta agttttataa gaaaaaaaag aatatataag
     6361 gcttttaaag cctttaaggt ttaacggttg tggacaacaa gccagggatg taacgcactg
     6421 agaagccctt agagcctctc aaagcaattt tgagtgacac aggaacactt aacggctgac
     6481 atggggcgcg cccag
//
